Davey M W, Sulkowski E, Carter W A
J Biol Chem. 1976 Dec 10;251(23):7620-5.
Interferons of human, mouse, and rabbit origin bind to straight chain hydrocarbons immobilized on agarose. The hydrophobic nature of binding is established by the following observations: (a) a positive correlation between the length of hydrocarbon ligand and the strength of interaction; (b) a stronger interaction with hydrocarbon ligands terminated with apolar rather than polar head groups; (c) a lack of dependence of binding on ionic strength and pH of the solvent; (d) a reversal of binding by ethylene glycol, a hydrophobic solute; (e) an increasing eluting efficacy of tetraalkylammonium ions with the length of their alkyl substituents. The hydrophobic interactions of human interferon underlie the efficiency of two-step chromatographic procedures. For example, human embryo kidney interferon can be purified about 3,600-fold by sequential chromatography on (a) concanavalin A-agarose, (b) octyl-agarose. Another two-step procedure: (a) concanavalin A-agarose, (b) L-tryptophan-agarose, gives about 10,000-fold purification. The overall recovery of interferon in both cases in close to 90%.
人、小鼠和兔源的干扰素可与固定在琼脂糖上的直链烃结合。结合的疏水性可通过以下观察结果得以证实：(a) 烃配体长度与相互作用强度之间呈正相关；(b) 与以非极性而非极性头基终止的烃配体相互作用更强；(c) 结合对溶剂的离子强度和pH值无依赖性；(d) 乙二醇（一种疏水性溶质）可使结合逆转；(e) 四烷基铵离子的洗脱效率随其烷基取代基长度的增加而提高。人干扰素的疏水相互作用是两步色谱法高效性的基础。例如，人胚肾干扰素可通过依次在(a) 伴刀豆球蛋白A - 琼脂糖、(b) 辛基 - 琼脂糖上进行色谱分离，纯化约3600倍。另一种两步法：(a) 伴刀豆球蛋白A - 琼脂糖、(b) L - 色氨酸 - 琼脂糖，可实现约10000倍的纯化。在这两种情况下，干扰素的总体回收率均接近90%。
